Biophysical Characterization of Interactions Involving Importin-α during Nuclear Import

Proteins containing the classical nuclear localization sequences (NLSs) are imported into the nucleus by the importin-α/β heterodimer. Importin-α contains the NLS binding site, whereas importin-β mediates the translocation through the nuclear pore. We characterized the interactions involving importin-α during nuclear import using a combination of biophysical techniques (biosensor, crystallography, sedimentation equilibrium, electrophoresis, and circular dichroism). Importin-α is shown to exist in a monomeric autoinhibited state (association with NLSs undetectable by biosensor). Association with importin-β (stoichiometry, 1:1; K D = 1.1 × 10 -8 M) increases the affinity for NLSs; the importin-α/β complex binds representative monopartite NLS (simian virus 40 large T-antigen) and bipartite NLS (nucleoplasmin) with affinities (K D = 3.5 × 10 -8 M and 4.8 × 10 -8 M, respectively) comparable with those of a truncated importin-α lacking the autoinhibitory domain (T-antigen NLS, K D = 1.7 × 10 -8 M; nucleoplasmin NLS, K D = 1.4 × 10 -8 M). The autoinhibitory domain (as a separate peptide) binds the truncated importin-α, and the crystal structure of the complex resembles the structure of full-length importin-α. Our results support the model of regulation of nuclear import mediated by the intrasteric autoregulatory sequence of importin-α and provide a quantitative description of the binding and regulatory steps during nuclear import.

Cellulose acetate membranes for osmosedimentation: Performance and morphological dependence on preparation conditions

Osmosedimentation is a new membrane-assisted separation technique, based on the rapid approach to sedimentation equilibrium when macromolecular solutions are contained within dialysis cells, in contact with solvent via a permselective membrane. Cellulose acetate membranes, cast from ternary solvent (acetone, acetic acid, water) solutions are suitable for osmosedimentation of proteins at low (2000 rpm) centrifugation speeds. Solute retention is improved when acetone-rich casting solutions are used. These membranes were examined by electron and optical microscopy, showing considerable morphological changes in the membrane support layer as the casting solution composition is changed. © 1986.

Sedimentation rates in Atibaia River basin, São Paulo State, Brazil, using Pb-210 as geochronometer

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

RECENT HEAVY-ION EXPERIMENTS AND STATISTICAL-MODEL OF HADRONS

The combined CERN and Brookhaven heavy ion (H.I.) data supports a scenario of hadron gas which is in chemical and thermal equilibrium at a temperature T of about 140 MeV. Using the Brown-Stachel-Welke model (which gives 150 MeV) we show that in this scenario, the hot nucleons have mass 3 pi T and the pi and rho mesons have masses close to pi T and 2 pi T, respectively. A simple model with pions and quarks supports the co-existence of two phases in these heavy ion experiments, suggesting a second order phase transition. The masses of the pion, rho and the nucleon are intriguingly close to the lattice screening masses.